ArticleMolecular and biochemical parasitology2017
CRISPR/Cas9 mediated sequential editing of genes critical for ookinete motility in Plasmodium yoelii.
Article in Molecular and biochemical parasitology, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 66 citations in OpenAlex.
- Histone H2B monoubiquitination drives sexual commitment in malaria parasites.Nature microbiology · 2026Article
- A divergent cyclic nucleotide binding protein promotes Plasmodium ookinete infection of the mosquito.PLoS pathogens · 2025Article
- Elevated NADNature communications · 2025Article
- Plasmodium yoelii as a model for malaria: insights into pathogenesis, drug resistance, and vaccine development.Molecular biology reports · 2025Review
- A scalable CRISPR-Cas9 gene editing system facilitates CRISPR screens in the malaria parasite Plasmodium berghei.Nucleic acids research · 2025Article
- Applications of CRISPR in Parasitology.Current pharmaceutical biotechnology · 2025Review
- A subpellicular microtubule dynein transport machinery regulates ookinete morphogenesis for mosquito transmission of Plasmodium yoelii.Nature communications · 2024Article
- An axonemal intron splicing program sustains Plasmodium male development.Nature communications · 2024Article
- Deaggregation of mutant Plasmodium yoelii de-ubiquitinase UBP1 alters MDR1 localization to confer multidrug resistance.Nature communications · 2024Article
- Large DNA fragment knock-in and sequential gene editing in Plasmodium falciparum: a preliminary study using suicide-rescue-based CRISPR/Cas9 system.Molecular and cellular biochemistry · 2024Article
- Plasmodium yoelii iron transporter PyDMT1 interacts with host ferritin and is required in full activity for malarial pathogenesis.BMC biology · 2023Article
- EB1 decoration of microtubule lattice facilitates spindle-kinetochore lateral attachment in Plasmodium male gametogenesis.Nature communications · 2023Article
- Exploring the Interactive Effects of Thymol and Thymoquinone: Moving towards an Enhanced Performance, Gross Margin, Immunity andAnimals : an open access journal from MDPI · 2022Article
- CRISPR/Cas9 and genetic screens in malaria parasites: small genomes, big impact.Biochemical Society transactions · 2022Review
- Recent Advances in Molecular Genetic Tools forVeterinary sciences · 2021Review
- A malaria parasite phospholipid flippase safeguards midgut traversal of ookinetes for mosquito transmission.Science advances · 2021Article
- Plasmodium transcription repressor AP2-O3 regulates sex-specific identity of gene expression in female gametocytes.EMBO reports · 2021Article
- Definition of constitutive and stage-enriched promoters in the rodent malaria parasite, Plasmodium yoelii.Malaria journal · 2020Article
- A protein palmitoylation cascade regulates microtubule cytoskeleton integrity in Plasmodium.The EMBO journal · 2020Article
- An intracellular membrane protein GEP1 regulates xanthurenic acid induced gametogenesis of malaria parasites.Nature communications · 2020Article
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Authors and funding
10 authors at 2 institutions in 2 countries.
Funding
Abstract
CRISPR/Cas9 has been successfully adapted for gene editing in malaria parasites including Plasmodium falciparum and Plasmodium yoelii. However, the reported methods were limited to editing one gene at a time. In practice, it is often desired to modify multiple genetic loci in a parasite genome. Here we describe a CRISPR/Cas9 mediated genome editing method that allows successive modification of more than one gene in the genome of P. yoelii using an improved single-vector system (pYCm) we developed previously. Drug resistant genes encoding human dihydrofolate reductase (hDHFR) and a yeast bifunctional protein (yFCU), with cytosine deaminase (CD) and uridyl phosphoribosyl transferase (UPRT) activities in the plasmid, allowed sequential positive (pyrimethamine, Pyr) and negative (5-fluorocytosine, 5FC) selections and generation of transgenic parasites free of the episomal plasmid after genetic modification. Using this system, we were able to efficiently tag a gene of interest (Pyp28) and subsequently disrupted two genes (Pyctrp and Pycdpk3) that are individually critical for ookinete motility. Disruption of the genes either eliminated (Pyctrp) or greatly reduced (Pycdpk3) ookinete forward motility in matrigel in vitro and completely blocked oocyst development in mosquito midgut. The method will greatly facilitate studies of parasite gene function, development, and disease pathogenesis.
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